Determination of fibrinogen content in cryoprecipitated antihemophilic factor by fluorescence quenching method
10.13303/j.cjbt.issn.1004-549x.2022.12.005
- VernacularTitle:荧光猝灭法测定冷沉淀凝血因子中的纤维蛋白原含量
- Author:
Jing LAN
1
;
Na FENG
1
;
Wenjuan ZHANG
1
;
Jin DING
1
;
Peng PENG
1
;
Na LIU
1
;
Xin CAO
1
;
Hua XU
1
Author Information
1. Shaanxi Blood Center, Xian 710000, China
- Publication Type:Journal Article
- Keywords:
fluorescein isothiocyanate labeled sheep anti-human fibrinogen;
fluorescence quenching;
cryoprecipitated antihemophilic factors;
fibrinogen content
- From:
Chinese Journal of Blood Transfusion
2022;35(12):1208-1212
- CountryChina
- Language:Chinese
-
Abstract:
【Objective】 To establish a new method for the determination of fibrinogen content in cryoprecipitated antihemophilic factor. 【Methods】 Fibrinogen (Fib) could bind with sheep anti-human fibrinogen (anti-Fib) specifically and further form antigen-antibody complex. When the Fib was present in the solution, the fluorescence of fluorescein isothiocyanate (FITC) labeled on the anti-Fib (FITC-anti-Fib) was quenched due to the formation of immune complex. The fluorescence quenching degree of FITC-anti-Fib was positively correlated with Fib concentration (cFib) in a certain concentration range. 【Results】 The linear relationship between fluorescence quenching degree [(I0-I)/I0] of FITC-anti-Fib and ln(cFib) was (I0-I)/I0=15.53ln(cFib)+ 80.79 (R2=0.99) when the cFib was in the range of (0.007 8-0.560 0) g/L. The recovery of Fib was (96.77-102.43) %. When the method was applied to determine Fib at high, medium, and low concentrations, the obtained intra-day variation coefficients were 0.31%, 0.56%, and 0.49%, respectively, and the inter-day variation coefficients were 3.81%, 3.06%, and 4.13%, respectively. There was no significant difference between the results measured by fluorescence quenching method and coagulation method (t=-0.075, P>0.05). 【Conclusion】 In this work, a new fluorescence method for the determination of Fib in cryoprecipitated antihemophilic factor was successfully established based on the specific combination of fib and FITC-anti-Fib. The method is simple and rapid. The obtained results were accurate and reliable by using this method to determine Fib.
